The effect of plant growth regulator on embryogenic tissue induction in four pine species

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Abstract

In this survey, the possibility of somatic embryogenesis in four different pine species was studied using different compositions of plant growth regulator. The embryogenic tissue induction phase was performed using mature zygotic embryo explant, culture medium DCR and using different levels of plant growth regulator in Pinus brutia, P. nigra, P. radiata and P. sylvestris species. The experiment was done through a completely randomized design with a simple factorial arrangement. Embryogenic tissue was measured in two phase, 30 and 40 days after the initial culture. After sterilizing of seeds kept at 4°C, and removing their coats, zygotic embryo was separated from megagametophyte and put into the Petri dish containing solid tissue culture. All the Petri dishes were maintained in darkness at 25°C. After 2 weeks the first signs of embryogenic tissue induction were observed. The results showed the significant effect of composition of plant growth regulator and specie on the amount of embryogenic tissue induction and the best result was obtained from P. brutia in hormon composition of 10µM 2,4-D+5µM BA. In order to proliferation of embryogenic tissues, subculture of tissues was performed every two weeks in new culture medium for six weeks. Finally, in order to maturation of somatic embryo from embryogenic tissue, the segments of embryogenic tissue with 5×5 mm dimensions were transferred to DCR culture medium. After two months, although quasi-embryo and abnormal structures got seen but the production of complete embryo from embryogenic tissue was unsuccessful and transition from preembryony phase and conversion.

Keywords

References

- Arya, S. Kalia, R. K. and Arya, I. D. 2000. Induction of somatic embryogenesis of Pinus  roxburghii
Sarg., Plant Cell Reports. 19: 775-780.
- Attree, S. M. Budimir, S. and Fowke, L. C. 1990. Somatic embryogenesis and plantlet regeneration from
cultured shoots and cotyledons of seedling from stored seed of black and white spruce (Picea mariana and
Picea glauca). Canadian Journal of Botany. 68(1): 30-34.
- Bozhkov, P. Filonova, L. H. and von Arnold, S. 2002. A key developmental switch during Norway
spruce somatic embryogenesis is induced by withdrawal of growth regulators and is associated with cell
death and extracellular acidification. Biotechnology and Bioengineering. 77(6): 658-667.
- Garin, E. Isabel, N. and Plourde, A. 1998. Screening of large numbers of seed families of Pinus strobus
L. for somatic embryogenesis from immature and mature zygotic embryos, Plant Cell Reports, 18: 37-43.
 - George, E. F. Michael, A. H. and De Klerk, G. J. 2008. Plant propagation by tissue culture, 3
 Ed.,Springer, Netherlands, 501 pp.
 - Gupta, P. K. and Durzan, D. J. 1985. Shoot multiplication from mature trees of Douglas fir (Pseudotsuga
menziesii) and sugar pine (Pinus lambertiana). Plant Cell Reports. 4: 177-180.
- Klimaszewska, K. Park, Y. S. Overton, C. Maceacheron, I. and Bonga, J. M. 2001. Optimized somatic
embryogenesis in Pinus strobus L., In Vitro Cellular & Developmental Biology-Plant, 37: 392-399.
- Lelu, M. A. Bastien, C. Drugeault, A. Gouez, M. L. and Klimaszewska, K. 1999. Somatic embryogenesis
and plantlet development in Pinus sylvestris and Pinus pinaster on medium with or without growth
regulators. Physiology Plant. 105: 719-728.
 - Isabel, N. and Tremblay, F. M. 1995. Somatic embryogenesis in red spruce (Picea rubens Sarg.), In: Jain,
S. Gupta, P. and Newton, R. (eds.), Somatic Embryogenesis in Woody Plants, Kluwer Academic,
Netherlands, 3: 111-123 pp.
- Maruyama, E. Hosoi, Y. and Ishii, K. 2007. Somatic embryogenesis and plantlet regeneration in
yakutanegoyon, Pinus armandii Franch. var. amamiana (Koidz.) Hatusima, an endemic and endangered
species in Japan, In Vitro Cellular & Developmental Biology-Plant, 43: 28-34 pp.
- Salajova, T. Salaj, J. and Kormutak, A. 1999. Initiation of embryogenic tissues and plantlet regeneration
from somatic embryos of Pinus nigra Arn. Plant Science. 145: 33-35.
- Stasolla, C. Kong, L. Yeung, E. C. and Thorpe, T. A. 2002. Maturation of somatic embryos in conifers:
morphogenesis, physiology, biochemistry and molecular biology, In Vitro Cellular & Developmental
Biology-Plant. 38: 93-105.
- Tang, W. and Newton, R. J. 2007. Micropropagation via organogenesis in slash pine, In: Jain, S. M. and
Häggman, H. (eds.), Protocols for Micropropagation of Woody Trees and Fruits, Springer, 15-22.
- Tautorus, T. E. Attree, S. M. Fowke, L. C. and Dunstan, D. I. 1990. Somatic embryogenesis from
immature and mature zygotic embryos and embryo regeneration from protoplasts in black spruce (Picea
mariana Mill). Plant Science. 67: 115-124.
- Tremblay, F. M. 1990. Somatic embryogenesis and plantlet regeneration from embryos isolated from
stored seed of Picea glauca. Canadian Journal of Botany. 68: 236-242.
- Wilson, S. M. and Thorpe, T. A. 1995. Somatic embryogenesis in Picea glauca (white spruce), P.
engelmannii  (engelmann spruce) and P. glauca engelmannii  complex (interior spruce), In: Jain, S. Gupta,
P. and Newton, R. (eds.), Somatic Embryogenesis in Woody Plants, Published by Kluwer Academic,
Netherlands, 3: 37-53.
 - Yildirim, T. Kaya, Z. and Isik, K. 2006. Induction of embryogenic tissue and maturation of somatic
embryos in Pinus brutia TEN., Plant Cell Tissue Organ Culture, 87: 67-76.
Forest and Wood Products
Volume 65, Issue 3 - Serial Number 3
October 2013
Pages 377-389

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